Boutsi E A
Department of Periodontology, Faculty of Dentistry, Tokyo Medical and Dental University.
Bull Tokyo Med Dent Univ. 1993 Mar;40(1):45-58.
The purpose of this study was to determine the immunodominant antigens of P. gingivalis 381 and to examine their composition and molecular weight. Fourteen periodontal patients, with high antibody titers to P. gingivalis 381 sonicated extract, were selected. Whole cell fraction, sonicated extract and outer membrane protein of P. gingivalis 381 were used as antigens in the untreated form as well as in the heated form and treated with papain. Total volume of sugar, protein and lipopolysaccharide (LPS) was estimated in each one of the three antigens. Antibody binding capacity to the three antigens was evaluated, before and after heat and papain treatment, by an enzyme-linked immunosorbent assay (ELISA). In addition, the immunoblotting analysis was performed. The quantitative assays showed that the whole cell fraction contained about ten times more LPS than the other two preparations while the outer membrane protein contained twice the amount of carbohydrates than the other two preparations. The 14 sera were classified into three groups according to the rate of reduction of antibody binding under heat and papain treatment. Concerning heat treatment, most of the sera showed a high reduction of antibody binding when reacting with the sonicated extract. However, antibody binding to the outer membrane protein antigen was hardly decreased by heat treatment in three sera. Also, these three sera showed almost the same response to the whole cell fraction antigen under heat treatment. Under papain treatment, almost all sera showed a moderate reduction of antibody binding when they reacted with the sonicated extract and whole cell fraction while they showed a low reduction of antibody binding when they reacted with the outer membrane protein. From the present study it could be concluded that a main proteinaceous antigen of P. gingivalis was recognized by the majority of the patients suggesting that the proteinaceous portion is an important part of the antigen, while some patients seemed to recognize the LPS or carbohydrate as the antigenic determinant.
本研究的目的是确定牙龈卟啉单胞菌381的免疫显性抗原，并检测其组成和分子量。选择了14名对牙龈卟啉单胞菌381超声提取物抗体滴度高的牙周炎患者。牙龈卟啉单胞菌381的全细胞组分、超声提取物和外膜蛋白以未处理形式以及加热形式用作抗原，并经木瓜蛋白酶处理。对三种抗原中的每一种估计糖、蛋白质和脂多糖（LPS）的总量。通过酶联免疫吸附测定（ELISA）评估热和木瓜蛋白酶处理前后对三种抗原的抗体结合能力。此外，进行了免疫印迹分析。定量分析表明，全细胞组分所含LPS比其他两种制剂多约十倍，而外膜蛋白所含碳水化合物量是其他两种制剂的两倍。根据热和木瓜蛋白酶处理下抗体结合的降低率，将14份血清分为三组。关于热处理，大多数血清在与超声提取物反应时显示出抗体结合的高度降低。然而，在三份血清中，热处理对外膜蛋白抗原的抗体结合几乎没有降低。同样，这三份血清在热处理下对全细胞组分抗原的反应几乎相同。在木瓜蛋白酶处理下，几乎所有血清在与超声提取物和全细胞组分反应时显示出抗体结合的适度降低，而在与外膜蛋白反应时显示出抗体结合的低降低。从本研究可以得出结论，大多数患者识别出牙龈卟啉单胞菌的一种主要蛋白质抗原，这表明蛋白质部分是抗原的重要组成部分，而一些患者似乎将LPS或碳水化合物识别为抗原决定簇。
